AS 3753—2001

AS 3753

Australian Standard™
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Recommended practice for chemical analysis by ultraviolet/visible spectrophotometry

This Australian Standard was prepared by Committee CH-016, Spectroscopy. It was approved on behalf of the Council of Standards Australia on 30 March 2001 and published on 5 May 2001.

The following interests are represented on Committee CH-016: Australian Chamber of Commerce and Industry Australian Government Analytical Laboratories
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CSIRO Energy Technology CSIRO Manufacturing Science and Technology Department of Defence Australia Department of Natural Resources, Queensland National Association of Testing Authority Queensland Health Scientific Services

Keeping Standards up-to-date Standards are living documents which reflect progress in science, technology and systems. To maintain their currency, all Standards are periodically reviewed, and new editions are published. Between editions, amendments may be issued. Standards may also be withdrawn. It is important that readers assure themselves they are using a current Standard, which should include any amendments which may have been published since the Standard was purchased. Detailed information about Standards can be found by visiting the Standards Australia web site at www.standards.com.au and looking up the relevant Standard in the on-line catalogue. Alternatively, the printed Catalogue provides information current at 1 January each year, and the monthly magazine, The Australian Standard, has a full listing of revisions and amendments published each month. We also welcome suggestions for improvement in our Standards, and especially encourage readers to notify us immediately of any apparent inaccuracies or ambiguities. Contact us via email at mail@standards.com.au, or write to the Chief Executive, Standards Australia International Ltd, GPO Box 5420, Sydney, NSW 2001.

This Standard was issued in draft form for comment as DR 00194.

AS 3753—2001

Australian Standard™
This is a free 6 page sample. Access the full version at http://infostore.saiglobal.com.

Recommended practice for chemical analysis by ultraviolet/visible spectrophotometry

Originated as AS CK19—1970. Previous edition AS 3753—1990. Second edition 2001.

COPYRIGHT
© Standards Australia International All rights are reserved. No part of this work may be reproduced or copied in any form or by any means, electronic or mechanical, including photocopying, without the written permission of the publisher. Published by Standards Australia International Ltd GPO Box 5420, Sydney, NSW 2001, Australia
ISBN 0 7337 3855 9

AS 3753—2001

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PREFACE
This Standard was prepared by the Standards Committee CH-016, Spectroscopy to supersede AS 3753—1990. The objective of this Standard is to describe recommended procedures, principles and techniques involved in the setting up and operation of UV/visible spectrophotometer for use in chemical methods for analysis. In preparing this revised Standard, the opportunity was taken to update the description of the components of UV/visible spectrophotometers in line with modern practice. Recognisance was also made of the variability of performance among the different instruments available in the market and in consequence, the manufacturer’s specifications for performance characteristics have been specified as the preferred criteria to be followed. The performance criteria specified in the superseded Standard have been retained as default criteria in cases where the manufacturer’s specifications are unavailable. The term ‘informative’ has been used in this Standard to define the application of the appendix to which it applies. An ‘informative’ appendix is only for information and guidance.

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3

AS 3753—2001

CONTENTS
Page SECTION 1 SCOPE AND GENERAL 1.1 SCOPE......................................................................................................................... 4 1.2 APPLICATION ........................................................................................................... 4 1.3 PRINCIPLE ................................................................................................................. 4 1.4 REFERENCED DOCUMENTS................................................................................... 4 1.5 DEFINITIONS............................................................................................................. 5 1.6 APPARATUS .............................................................................................................. 5 1.7 SAFETY PRECAUTIONS .......................................................................................... 5
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SECTION 2 INSTRUMENTATION 2.1 GENERAL................................................................................................................... 6 2.2 SOURCES OF RADIATION....................................................................................... 6 2.3 WAVELENGTH SELECTION.................................................................................... 6 2.4 OPTICAL SYSTEM .................................................................................................... 7 2.5 MEASUREMENT SYSTEM ....................................................................................... 7 2.6 SAMPLE INTRODUCTION ....................................................................................... 9 SECTION 3 OPTIMIZATION 3.1 INTRODUCTION ..................................................................................................... 11 3.2 CELL CARE AND HANDLING ............................................................................... 11 3.3 MONITORING SPECTROPHOTOMETER FUNCTIONS ....................................... 13 3.4 FACTORS AFFECTING ABSORPTION.................................................................. 17 3.5 BACKGROUND ABSORPTION .............................................................................. 17 3.6 MATRIX EFFECTS .................................................................................................. 18 3.7 SETTING UP THE INSTRUMENT .......................................................................... 18 3.8 ADDITIONAL CONSIDERATIONS ........................................................................ 19 SECTION 4 CALIBRATION AND ANALYSIS 4.1 REAGENT PURITY.................................................................................................. 20 4.2 REFERENCE MATERIALS ..................................................................................... 20 4.3 IDENTIFICATION USING WAVELENGTH SCANNING...................................... 20 4.4 DETERMINATIONS AT FIXED WAVELENGTHS................................................ 21 4.5 CALIBRATION AND TEST PROCEDURE............................................................. 22 4.6 ACCEPTANCE OF RESULTS.................................................................................. 22 APPENDICES A DEFINITIONS OF TERMS USED IN ULTRAVIOLET/VISIBLE SPECTROPHOTOMETRY ....................................................................................... 24 B FLOWSHEET ON THE PROCEDURE FOR THE ACCEPTANCE OF ANALYTICAL VALUES FOR TEST SAMPLES .................................................... 28

AS 3753—2001

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STANDARDS AUSTRALIA Australian Standard Recommended practice for chemical analysis by ultraviolet/visible spectrophotometry

S EC TION
1.1 SCOPE
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1

S C OP E

AND

G E NER A L

This Standard sets out recommendations for instrumentation and operating techniques suitable for chemical analysis of solutions by ultraviolet/visible spectrophotometry, and includes a summary of testing procedures, calibration procedures and recommendations for safe operation. Although most sections of this Standard refer to the analysis of solutions, sections on instrumentation and instrument optimization are relevant to instruments used for gas analysis. 1.2 APPLICATION This Standard is intended to be read in conjunction with the instrument manufacturer’s recommendations. 1.3 PRINCIPLE Ultraviolet/visible spectrophotometry relies upon the following: (a) The absorption of radiation by a substance where the wavelength of the radiation corresponds to the energy required to raise the substance to a state of higher energy (excited state) in an allowed electronic transition. Absorption may be highly wavelength specific or may occur over a broad wavelength range. The excited state of the substance being unstable and reverting rapidly to the normal state without emission of radiation. The absorbance by the substance being a function of the number of absorbing molecules in the light path (see Appendix A, Bouguer’s (Lambert’s) Law and Beer’s Law).

(b) (c)

The technique described in this Standard involves passing radiation of known wavelength through a solution of a sample, and determining the absorbance at that wavelength. 1.4 REFERENCED DOCUMENTS The following documents are referred to in this Standard: AS 2162 2162.1 2162.2 2164 2166 2167 Verification and use of volumetric apparatus Part 1: General—Volumetric glassware Part 2: Guide to the use of piston-operated volumetric apparatus (POVA) Laboratory glassware—One-mark volumetric flasks One-mark pipettes Graduate straight pipettes

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AS 3753-2001, Recommended practice for chemical analysis by ultraviolet/visible spectrophotometry

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